Analysis of changes in the percentage of B (CD19) and T (CD3) lymphocytes,subsets CD4, CD8 and their memory (CD45RO), and naive (CD45RA) T cells in children with immune and non-immune thyroid diseases

Graves' disease (GD) is an autoimmune thyroid disease caused by immunological abnormality. The immune cells (lymphocytes T and B) which infiltrate the thyroid gland play a key role in the development of autoimmune thyroid disease (AITD). The aim of this study was to evaluate the differences between distribution of T (CD3) lymphocytes, subsets CD4, CD8, and their memory (CD45RO), and naive (CD45RA) T cells and B (CD19) lymphocytes in the peripheral blood of patients with Graves' disease (GD) (n = 33, mean age 15.9 +/- 5.9 years) and non-toxic nodular goiter (NTNG) (n = 25, mean age 15.2 years), in comparison to age- and sexmatched healthy control subjects (n = 25, mean age 15.9 years). The percentages of peripheral blood lymphocyte subsets were analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. In the untreated Graves' patients we observed an increase in the percentage of CD19+ (p<0.007, p<0.003), CD4+ (p<0.004, p<0.017), CD4+CD45RO+ (p<0.04, NS), CD4/CD8 ratio (p<0.002, p<0.001) and a decrease in the percentage of CD8+ (p<0.02, p<0.02), CD4+CD45RA+ (p<0.04, p<0.03) cells in comparison to the healthy control subjects and euthyroid Graves' patients. These abnormalities were absent in children with non-toxic nodular goiter. In addition, the levels of CD3+, CD4+CD8+, CD8+CD45RO+ T cells and CD8 lymphocytes co-expressing CD45RA and CD45RO antigens were similar in all groups and no statistically significant differences were found in comparison to the healthy controls. In the untreated Graves' patients we found a positive correlation between serum levels of fT4 and fT3 and the percentage of CD19+ lymphocytes (r = 0.45, p<0.01, r = 0.37, p<0.04), between serum level of fT4 and the percentage of CD4CD45RO (r = 0.4, p<0.02) lymphocytes and between concentration of TRAb and CD4+ (r = 0.38, p <0.04) and CD19+ (r = 0.39, p<0.016) cells. Statistically significant negative correlations existed between TRAb, TPO-Ab or TG-Ab concentration in blood serum and the percentage of CD8+ lymphocytes (r = -0.55, p<0.002; r = -0.41, p<0.02; r = -0.51, p<0.004), and between fT4 concentration and the percentage of CD8+ (r = -0.39, p<0.02) lymphocytes. No such correlation was detected in patients with non-toxic nodular goiter. We conclude that the abnormal distribution of B lymphocytes, memory and naive T cell subsets in the peripheral blood in children and adolescents with untreated Graves' disease suggests their role in the development of autoimmunity. The normalization in the percentage of these immune cells after thyrostatic treatment in comparison to newly diagnosed patients confirms the immunomodulatory effect of methimazole therapy.     

RSV感染患儿外周血CD_4~*”CD_(45)RO~+CD_(45)RA~+表达变化的研究

目的：探讨下呼吸道感染呼吸道合胞病毒(RSV)患儿外周血辅助性T淋巴细胞(CD4),原始T细胞(CD45RA+),记忆性T细胞(CD45RO+)表达的变化。方法：用单克隆抗体免疫荧光标记,流式细胞仪检测30例RSV下呼吸道感染患儿急性期外周血单个核细胞(PBMCs)CD4+,CD45RA+细胞,其中11例同时检测CD45RO+细胞,同期检测9例年龄、性别无差异的健康儿为对照。结果：RSV下呼吸道感染组患儿CD4为(32.74±10.60)%,明显低于对照组(40.76±6.82)%,2组有显著性差异(P0.05)。结论：RSV感染急性期存在免疫功能紊乱,外周血CD4,CD45RO+下降,而CD45RA+明显增加,这可能是CD45RO+向呼吸道迁移的结果。     

Cytokine production, activation marker, and skin homing receptor in children with atopic dermatitis and bronchial asthma

T cells are known to develop a critical role in the pathogenesis of atopic dermatitis (AD) and bronchial asthma. T cells involved in AD express the skin homing receptor CLA, but no lung homing receptor has been identified in bronchial asthma. We compared different cell markers and the cytokine production in T cells from children with AD or bronchial asthma. We studied the involvement of CLA+ and CLA- T-cell subpopulations in these diseases. We studied 20 children with acute AD lesions, 15 with mild persistent asthma, and 15 non-atopic controls. All patients were sensitized to house dust mite (DP) and evaluated during the acute phase. Total and specific IgE were measured by immunoassay and the expression of different cell markers and the cytokine production was analyzed by flow cytometry in peripheral blood mononuclear cells. Total IgE was significantly higher in AD children and IgE to DP in the asthmatic children. There was a significant increase in CD25+ CD4+ cells in asthmatic children and in HLA-DR+ CD4+ and HLA-DR+ CD8+ cells in AD. In the CD4+ subsets, there was an increase in IL-13, IL-5 and TNF-alpha in AD compared to controls, a decrease in IFN-gamma in asthmatic children compared to controls, and an increase in IL-13, IL5, IL2, TNF-alpha, and IFN-gamma in the AD compared to asthmatic children. Changes in cytokine production were mainly detected in CLA+ cells in AD and in CLA- cells in asthma. Differences exist in total and specific IgE, activation markers, and cytokine patterns between AD children and children with asthma, with the former expressing a Th2 pattern whereas in asthmatic children we only detected a decrease in IFN-gamma. Moreover, the subpopulations (CLA+ vs. CLA-) expressing these changes were different, indicating that the underlying mechanisms in the two diseases are not exactly the same.     

CD95 (APO-1/Fas) expression on naive CD4(+) T cells increases with disease progression in HIV-infected children and adolescents: effect of highly active antiretroviral therapy (HAART)

We studied the expression of the CD95 receptor (APO-1/Fas) on peripheral blood T cell subpopulations in 37 HIV-1-infected children and adolescents stratified according to disease stage or antiretroviral treatment regimen and compared the results to values obtained in 12 healthy age-matched control subjects. CD95 expression on CD45RA(+) CD45RO(-)/CD62L(+) (resting/naive) and CD45RO(+) CD45RA(-) (primed/memory) CD4(+) and CD8(+) T cells was assessed quantitatively by four-color and three-color flow cytometry. CD4(+) T cells contained a population of predominantly CD95(-) resting/naive cells and a population of CD95(high) primed/memory cells, whereas CD8(+) T cells had a more uniform pattern of CD95 expression. The percentage of CD95(+) CD4(+) T cells increased with disease progression because of both an augmented median fluorescence intensity on resting/na?ve cells and an increased percentage of CD95(high) cells. Patients with highly active antiretroviral combination therapy who maintained stable CD4 counts in the presence of elevated plasma viral load had nearly normal numbers of CD95(-) resting/naive CD4(+) T cells, whereas CD95 expression in the CD8(+) T cell subset was still elevated compared with control subjects. Low CD95 expression on resting/naive CD4(+) T cells may therefore indicate a low risk for disease progression in antiretrovirally treated and untreated patients.     

肺炎支原体感染患儿T淋巴细胞亚群检测及分析

目的　观察肺炎支原体肺炎患儿急性期外周血T淋巴细胞亚群、T淋巴细胞活化状态的改变,探讨其发病机制。方法　采用流式细胞仪技术检测了2 0 0 2年1 0月至2 0 0 3年6月就诊于上海市金山区中心医院的1 7例肺炎支原体肺炎患儿急性期外周血T淋巴细胞亚群及T细胞亚群上CD2 5+的表达和CD4+细胞上CD4+CD45RA+、CD4+CD45RO+的表达;对照组为1 0例健康体检儿童。两组年龄、性别差异无显著意义。结果　肺炎支原体肺炎患儿急性期外周血CD3 +百分率( % )为( 62 . 2 3±6 .2 7) ,较对照组( 68 .60±4. 74)低,差异有显著性意义(P <0 . 0 5) ;CD4+、CD8+百分率较对照组差异无显著性意义(P >0. 0 5) ;CD8+CD2 5+百分率( % )为( 0 . 61±0 . 58) ,较对照组( 2 .1 6±0 . 40 )降低,差异有极显著性意义(P <0 .0 1 ) ;CD4+CD45RA+/CD4+CD45RO+比值与对照组相比降低(P <0 . 0 5)。结论　肺炎支原体肺炎时存在细胞免疫失调,主要表现为总T细胞降低,T细胞活化障碍和CD4+CD45RA+/CD4+CD45RO+平衡失调。     

Age-related changes in intracellular TH1/TH2 cytokine production, immunoproliferative T lymphocyte response and natural killer cell activity in newborns, children and adults

To evaluate the development of the neonatal immune system, we measured T lymphocyte response to Con A, intracellular IL-2, IL-4, IFN-gamma and IL-10 production, and natural killer cell (NKC) activity in 12 very preterm, 12 preterm and 20 term neonates, 10 children and 10 adults. Immunoproliferation to Con A was significantly lower in cord blood than in children or adults. The percentage of CD4+ lymphocytes was significantly higher in newborns while CD8+ cells were higher at older ages, with a resulting gradual decline of the CD4+/CD8+ ratio. The percentage of IL-2-producing CD4+ and CD8+ cells was higher in all newborn groups than in children and adults, while the percentage of IL-4-producing cells was higher for CD8+ and lower for CD4+ cells in cord blood than in children and adults. Neonates had substantially lower percentages of CD4+ and CD8+ IFN-gamma-producing cells. A significant negative correlation was observed between gestational age and IFN-gamma-CD4+-, IL-2-CD8+-, and IL-10- CD4+-producing cells. In addition, a positive correlation was found between gestational age and IL-10-CD8+-producing cells. Percentages of CD4+/CD45RA+ cells were higher and CD4+/CD45RO+ percentages were lower in newborns than in children and adults. NKC activity in infants was significantly correlated with gestational age and significantly impaired compared to children and adults. On the whole, these results suggest a gradual development of immunity during gestation and show significant immaturity of cellular immune response at birth. The reduction of NKC activity, the lower proliferative response of T cells, the reduced cytotoxic response and a dysregulated cytokine production may contribute to the neonatal increased risk of infection and to the low incidence of graft-versus-host disease after cord blood transplantation.     

Immunophenotypic characterisation of peripheral blood lymphocytes in autoimmune polyglandular syndrome type 1: clinical study and review of the literature

Autoimmune endocrinopathies are characterised by an increased number of peripheral blood lymphocytes (PBL) expressing activation/ memory markers on their surface. The aim of this study was to determine whether a similar finding could be detected in a group of 11 paediatric and young adult patients suffering from autoimmune polyglandular syndrome type 1 (APS1), also called autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), as very few data have previously been reported in this field. The control group was made up of 11 sex- and age-matched healthy subjects. Fifteen lymphocyte subsets were compared, in terms of percentage and absolute number, and statistical analysis was performed by the Mann-Whitney test. Measurement of T (CD3+), B (CD19+), natural killer (NK, CD3-CD16/56+), CD4+ and CD8+ T lymphocytes showed that patients with APS1 had a higher percentage and absolute count of T lymphocytes: this was entirely due to the statistically larger CD3+CD4+ fraction. Patients with APS1 also had slightly fewer B and NK lymphocytes, but the difference was negligible. Comparison of CD4+ subpopulations bearing activation and naive/memory antigens (marked by CD69, CD25, anti-HLA-DR, CD45RA and CD45RO) showed that patients with APS1 had generally larger percentages and absolute counts of these subsets: however, only the percentage and absolute size of the CD4+CD25+ subset (p = 0.0354 and p = 0.0151, respectively), and the absolute number of the CD4+ anti-HLA-DR+ and CD4+ CD45RO+ subsets (p = 0.0193 and p = 0.0209, respectively) were significantly higher. Interestingly, patients with APS1 also had significantly fewer CD8+CD11b+ and CD3-CD8+ cells. In conclusion, PBL distribution in APS1 resembles that of other autoimmune diseases. Further studies are needed to confirm and possibly extend these data.     

Lymphocyte subsets in human tonsils: The effect of age and infection

The aim of the present study was to determine the effect of repeated tonsillitis on the development of lymphocyte subsets in the tonsils and among peripheral blood lymphocytes (PBL) of children. Subsets of T- and B cells were analyzed in the tonsils and in PBL of patients undergoing tonsillecto-my for idiopathic tonsillar hypertrophy, recurrent tonsillitis, or tonsillar hypertrophy and tonsillitis. The majority of the CD4+ cells in the tonsils displayed the CD45RO+ phenotype, while the majority of those in the PBL displayed the CD45RA+ phenotype. Likewise, the proportion of CD45RO+CD8+ cells was higher in the tonsils than among PBL. The proportion of CD4 cells expressing the CD45RO marker increased with age among PBL, but not in the tonsils. B cells, detected by their CD 19, CD20, and CD21 markers, were three times more abundant in the tonsils than in the PBL. The proportion of CD38+ cells showed a negative correlation with age, both in the tonsils and among PBL. Among PBL a striking age-related reduction was seen in the proportion of CD19+, CD21+ and CD38+CD21+ B cells. In contrast, in the tonsils age-related changes could be detected only in the proportion of CD21+CD38+ cells. No difference among patients with various clinical diagnoses was detectable in any of the T- and B cell subsets in the tonsils and PBL. Thus, lymphocyte subsets evolve independently in the tonsils and peripheral blood, with the repeated antigenic challenge of tonsillar lymphocytes not influencing circulating memory cells.     

儿童原发性免疫缺陷病的免疫学改变及分析

目的了解儿童原发性免疫缺陷病(PID)患儿的实验室免疫检查特点,为临床早期诊断PID提供依据。方法对76例PID患儿的免疫学检查包括细胞免疫和体液免疫指标进行回顾性总结及分析。结果体液免疫检查中,普通变异型免疫缺陷病(CVID)及联合免疫缺陷病的IgG均降低,尤以CVID降低更为明显,且伴IgA及CD19+B细胞明显降低;IgA降低主要见于CVID、选择性IgA缺乏症(IgAD)及共济失调毛细血管扩张症(AT);低丙种球蛋白血症患儿IgG、IgA及CD19+B细胞均降低。细胞免疫功能中CD3+、CD4+T细胞降低主要见于细胞免疫缺陷病、联合免疫缺陷病及DiGeorge综合征(DGS);AT患儿的CD4+/CD8+比例也降低。8例CVID患儿检测T细胞功能亚群及活化指标,其中5例患儿的CD4+/CD8+比例倒置(CD4+/CD8+<0.9);与对照组相比,CVID患儿的CD4+HLA-DR+和CD8+CD45RO+T细胞比例明显升高,CD4+CD45RA+T细胞比例明显降低(P均<0.01)。结论儿童原发性免疫缺陷病免疫学改变各异。CVID患者体内T细胞过度活化,可能是CVID患者容易罹患自身免疫性疾病的原因之一。     

High CD4/CD45RO+ and CD8/CD45RO+ frequencies in children with vaccine-modified measles

BACKGROUND: Despite availability and wide vaccine coverage, measles infections still occur especially in developing countries. An outbreak of measles occurred among previously immunized older Ghanaian children who had milder clinical symptoms with measles-specific IgG antibodies that could have been attributed to secondary vaccine failure, suggesting that the infection was vaccine-modified measles (VMM). METHODS: Two-color immunophenotyping of the peripheral blood mononuclear cells was performed at acute, recovery and convalescence phases for 19 VMM patients (mean age 6.2 +/- 3.5 years) using flow cytometry, and compared with that of 20 healthy, sex- and age-matched controls. RESULTS: The results showed a significantly higher memory helper (CD4(+)/CD45RO(+)) cell frequency and increased suppressor cell (CD8(+)/CD45R0(+)) frequency in VMM patients compared to healthy controls. There were no complications and all the patients recovered completely. CONCLUSIONS: These findings show that the mild symptoms in patients with VMM may have correlated with the increase of memory T cells, which is in sharp contrast with previous reports on acute measles infection. This may suggest that the intact immunologic memory cells could have been crucial for the resolution of VMM.     

Activated peripheral blood CD4 and CD8 T-lymphocytes in child asthma: correlation with eosinophilia and disease severity

There now exists compelling evidence of a role for cell-mediated immunity in the pathogenesis of adult asthma, but little information is available as to what extent this process participates in the pathogenesis of childhood asthma. We hypothesised that asthma in children is associated with the activation of T-lymphocytes whose products regulate, at least in part, the mobilisation and recruitment of eosinophils and thereby disease severity. Our aims, therefore, were to compare the expression of activation markers, including CD45 isoforms, on peripheral blood T-lymphocytes from asthmatic and non-asthmatic, allergic control children matched for age and atopic status, and to attempt to correlate the percentages of activated T-lymphocytes in the asthmatics with the numbers of peripheral blood eosinophils and with disease severity. Seventeen children with moderate to severe chronic asthma were compared with 8 non-asthmatic, allergic children matched for age and atopic status. Expression of the activation markers CD25, HLA-DR and VLA-1 and the CD45 isoforms CD45RA and CD45RO on peripheral blood CD4 and CD8 T-lymphocytes was measured using dual fluorescence flow cytometry. Peripheral blood eosinophils were measured using an automated laser cytometer. Asthma severity was assessed by a symptom score, spirometry and measurement of histamine PC₂₀. The absolute numbers of eosinophils in the peripheral blood of the asthmatics were elevated as compared to the non-asthmatic, allergic controls (p < 0.01), whereas the absolute numbers of both CD4⁺ and CD8⁺ T-lymphocytes were not significantly different. The percentages and absolute numbers of CD4⁺ T-lymphocytes expressing CD25 and HLA-DR and CD8⁺ T-lymphocytes expressing CD25 also were significantly elevated in the asthmatics as compared with the controls (p < 0.04 in each case). However, the percentages and absolute numbers of CD4⁺ T-lymphocytes expressing VLA-1 and CD8⁺ T-lymphocytes expressing HLA-DR and VLA-1 did not significantly differ between the asthmatics and controls. Similarly, the percentages and absolute numbers of both CD4⁺ and CD8⁺ T-lymphocytes expressing the CD45 isoforms CD45RA and CD45RO were not significantly different in the asthmatics and controls. In the asthmatics, the numbers of peripheral blood eosinophils correlated with disease severity as measured by histamine PC₂₀ (p = 0.02). The percentages of CD4⁺ T-lymphocytes expressing HLA-DR correlated both with the numbers of peripheral blood eosinophils and with disease severity (histamine PC₂₀ (p < 0.03)). In addition, the percentages of CD8⁺ T-lymphocytes expressing CD25 correlated positively with disease severity as measured by a symptom and therapy score (p = 0.03). Finally, the percentages of CD8⁺ T-lymphocytes expressing HLA-DR correlated with histamine PC₂₀ (p = 0.03). These observations are consistent with the hypothesis that activated T-lymphocytes play a role in the pathogenesis of childhood asthma at least in part through their influence on eosinophil recruitment.     

CD+8 CD-28调节性T细胞在传染性单核细胞增多症中的作用初探

目的观察传染性单核细胞增多症（IM）患儿CD8^＋CD28^-调节性T（Tr）细胞动态改变,探讨急性EB病毒感染的免疫调控机制。方法观察25例IM患儿及相同数量同龄对照组。用流式细胞术检测外周血CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD8^＋CD28^＋表面标志;用逆转录-聚合酶链反应和实时定量聚合酶链反应检测CD8^＋CD28^-Tr细胞中IL-6、IL-10、IFN—γ及MC中ILT-3、ILT-4mRMA表达。结果IM患儿急性期CD8^＋CD28^-Tr细胞阳性表达率明显高于同龄对照组（P〈0．01）,恢复期阳性表达率较急性期有所下降,但仍高于对照组（P〈0．01）;IM患儿CD8^＋CD28^-Tr细胞和MC效应分子IL-6、IL-10、IFN-γ、ILT-3、ILT-4表达明显增高（P〈0．01）。结论IM患儿CD8^＋CD28^-Tr细胞增多可能是机体调控病毒感染的适应性免疫反应过程,其数量（及活性）降低或增高是否与EB病毒感染相关性疾病的发生发展有关尚需进一步探讨。     

Prenatal exposure to house dust mite allergen (Der p 1), cord blood T cell phenotype and cytokine production and atopic dermatitis during the first year of life

This study investigated the influence of prenatal exposure to house dust mite (HDM, D. pteronyssinus) on interleukin (IL)-2, interferon-gamma (IFN-gamma) and IL-4 producing CD4(+) and CD8(+) T lymphocytes in cord blood as well as on the development of sensitization and occurrence of atopic dermatitis (AD) as the first symptom of allergy during the first year of life. Dust samples (n = 22) were collected by vacuum cleaning the maternal mattress during early to mid-pregnancy. In these samples, the amount of the major HDM antigen (Der p 1) was assessed by a sensitive enzyme-linked immunosorbent assay technique (detection limit 0.004 microg/g dust). Flow cytometry was used to determine cord blood lymphocyte subtypes and to quantify the intracellular amounts of IL-2, IFN-gamma and IL-4 produced by cord blood CD4(+) helper and CD8(+) cytotoxic T lymphocytes, both spontaneously and after stimulation with phorbol-12-mirystate-13-acetate and ionomycin. Children were followed for 1 yr for the presence of symptoms associated with allergy. In addition, at the age of 1 yr specific IgE to different classical inhalant and food allergens was measured. Higher prenatal exposure to Der p 1 (>0.2 microg/g dust) was associated with a significant lower percentage of IFN-gamma producing stimulated CD4(+) T lymphocytes, compared with lower prenatal Der p 1 exposure (p = 0.03). The presence of AD during the first year of life (n = 9) was associated with an increased number of naive CD4(+) CD45RA(+) lymphocytes (p = 0.03), with an increased spontaneous IL-4 production by CD8(+) lymphocytes (p = 0.04) and with a decreased percentage of IFN-gamma producing stimulated CD4(+) lymphocytes (p = 0.04). Furthermore, exposure to HDM during pregnancy tended to be higher in mothers of children with AD during the first year of life when compared with those without AD (p = 0.08). This study shows that the level of prenatal exposure to Der p 1 influences the immune profile of cord blood T lymphocytes and the clinical outcome in early life. Therefore, the prenatal environment must be regarded as a possible early risk factors for allergic diseases in children.     

����������ԭ���������ۺ����Ի���CD4+CD25+������Tϸ����Ӱ�����

目的 观察原发性肾病综合征(PNS)患儿泼尼松治疗前后CD4+CD25+调节性T细胞(CD4+CD25+Tr)的变化,阐明肾上腺糖皮质激素治疗儿童PNS的免疫机制.方法 选择2004-2007年在深圳市儿童医院住院治疗的初发PNS患)L42例,其中激素敏感型32例,激素耐药型10例.同期25例健康体检儿童作为对照组.流式细胞术检测泼尼松治疗前后外周血CD3+CD4+CD8-、CD3+CD4-CD8+、CD4+CD25+Tr的比例,荧光定量聚合酶链反应(Real-time PCR)检测泼尼松治疗前后外周血单个核细胞(PBMC)叉头型基因P3(Foxp3)、细胞毒性T淋巴细胞相关抗原4(CTL-4)和糖皮质激素诱导的肿瘤坏死因子受体(GITR)基因mRNA的表达.结果 与对照组比较,PNS患儿外周血CD3+CD4+CD8-T细胞、CD3+CD4-CD8+T细胞、CD4+CD25+Tr比例无明显改变(P>0.05).激素敏感型PNS患儿CD4+CD25+Tr比例在泼尼松治疗后明显增高,差异有统计学意义(P<0.01);激素耐药型PNS患儿CD4+CD25+Tr比例在泼尼松治疗前后无明显改变(P>0.05).激素敏感型PNS患儿在泼尼松治疗后PBMC细胞Foxp3、CTLA-4和GITR基因mRNA的表达明显增高;而激素耐药型PNS患儿泼尼松治疗前后Foxp3、CTLA-4基因表达无明显改变,仅GITR表达明显增高.结论 泼尼松等肾上腺糖皮质激素类药物可通过上调激素敏感型PNS患儿CD4+CD25+调节性T细胞的表达发挥免疫治疗作用.     

CD4+CD25int/highCD127low 调节性T细胞在再生障碍性贫血患儿中的检测及意义

目的：探讨再生障碍性贫血（AA）患儿外周血 CD4+ CD25int/high CD127low 调节性 T 细胞（Treg）的表达特点及其与 Hb、WBC 和 血小板（Plt）数量的相关性。方法：采用流式细胞术检测 15 例正常儿童（对照组）和 22 例治疗有效的AA患儿治疗前后外周血 CD4+ CD25int/high CD127low Treg百分比的变化,分析 CD4+CD25high-CD127low Treg与 Hb、WBC 和 Plt 数量的相关性。结果：AA 组急性期外周血 CD4+CD25+、CD4+CD25high、CD4+CD25+CD127low、CD4+CD25highCD127low Treg占 CD4+ T 细胞的百分比均较对照组显著降低（P﹤0.05）;恢复期的表达则高于急性期（P﹤0.05）,而与对照组比较差异无统计学意义（P﹥0.05）。AA组急性期、恢复期及对照组的 CD4+CD25highCD127low Treg表达率与外周血中 Hb、WBC 和 Plt 数量均呈正相关（r分别为 0.499、0.526、0.540,P<0.05）。结论：CD4+CD25int/highCD127low Treg 减低可能与 AA 发病有关,为其作为判断 AA 病情变化的指标提供了可能的理论依据,并可以作为免疫治疗的靶点之一进行深入研究。     

幼年特发性关节炎患儿外周血Th细胞亚群变化

目的探讨幼年特发性关节炎(JIA)患儿外周血CD4 T淋巴细胞及其亚群CD4 CD45RA 、CD4 CD45RO 的表达及其临床意义。方法采用免疫荧光标记技术和流式细胞仪检测36例JIA患儿外周血CD4 T淋巴细胞及其亚群CD4 CD45RA 、CD4 CD45RO 的表达,同期检测20例年龄、性别无差异的健康儿童为对照。结果JIA患儿外周血CD4 T淋巴细胞明显低于对照组(t=2.099,P<0.05),CD4 CD45RA T淋巴细胞数与对照组比较明显降低(t=3.450,P<0.01),CD4 CD45RO T淋巴细胞数明显升高(t=3.913,P<0.01),CD4 CD45RA T/CD4 CD45RO T比值明显降低(t=4.904,P<0.01);与对照组比较,JIA各亚型(全身型、多关节型、少关节型)的CD4 CD45RO T淋巴细胞数均明显升高,CD4 CD45RA T/CD4 CD45RO T比值明显降低(P<0.01);CD4 CD45RA T淋巴细胞数与正常对照组比较在全身型中显著降低(t=4.192,P<0.01);在多关节型中明显降低(t=2.214,P<0.05);在少关节型中稍有降低,但与对照组比较差异无显著性(t=1.793,P>0.05)。结论JIA患儿的免疫功能紊乱主要表现为CD4 T淋巴细胞及其亚群CD4 CD45RA T/CD4 CD45RO T失衡,这可能在JIA发病机制中起着重要的作用。     

高效抗逆转录病毒治疗对艾滋病患儿CD8+T细胞活化分子CD38和人类白细胞抗原DR表达水平的影响及其与病毒载量的关系

目的 观察高效抗逆转录病毒治疗(HAART)对我国艾滋病患儿CD8+T细胞活化标志分子CD38和人类白细胞抗原DR(HLA-DR)表达水平的影响及其与病毒载量的关系.方法 对194例接受HAART的艾滋病患儿进行横断面研究,用流式细胞术检测CD4+、CD8+T细胞数,以及CD8+/CD38+和CD8+/HLA-DR+T细胞比例,RT-PCR检测血浆HIV RNA载量.并检测52名健康儿童CD8+/CD38+和CD8+/HLA-DR+T细胞比例作为正常对照.结果 本组中,135例病毒载量＜400 copies/ml,59例病毒载量≥400 copies/ml.病毒载量≥400 copies/ml组CD8+/CD38+T和CD8+/HLA-DR+T细胞水平显著高于病毒载量＜400 copies/ml组,差异有统计学意义(29.6±10.1 vs19.9±9.8;17.7±6.4 vs 9.6±6.1,P＜0.05);病毒载量＜400 copies/ml组,CD8+/CD38+T细胞接近正常水平,而CD8+/HLA-DR+T细胞仍高于正常水平,差异有统计学意义(19.9±9.8 vs 15.6±9.0;9.6±6.1 vs 5.8±3.3,P＜0.05).CD8+/CD38+和CD8+/HLA-DR+T细胞百分比均与病毒载量成正相关(前者相关系数R=0.403,P=0.03,后者相关系数R=0.569,P=0.09).结论 艾滋病患儿CD8+T细胞活化程度与病毒载量成正比,有效的HAART治疗能够显著地降低HIV感染者免疫活化程度,CD8+/CD38+和CD8+/HLA-DR+T细胞百分比可能是资源有限地区替代病毒载量检测的潜在指标.

Abstract：

Objective To study the expression of CD38 and HLA-DR on CD8 + T cells in pediatric AIDS patients receiving highly active antiretroviral therapy (HAART) and the relationship of immune activation and disease progression. Methods A cross-section study of 194 pediatric AIDS patients receiving HAART was carried out and 52 age-matched healthy children were recruited as control. The percentage of CD4+ , CD8+ , CD8+/CD38 + and CD8+/HLA-DR+ T cells was tested using flow cytometry, and HIVRNA in plasma was detected by quantitative RT-PCR. Results One hundred and ninety-four pediatric AIDS patients were divided into two groups according to the viral load: 59 patients with VL≥400 copies/ml and 135 patients with VL＜400 copies/ml. The percentage of CD8 +/CD38+ and CD8 +/HLA-DR+ T cells of patients with VL≥400 copies/ml was significantly higher than that of patients with VL ＜ 400 copies/ml (P ＜ 0. 05 ). Of patients with VL ＜ 400 copies/ml, the percentage of CD8 +/CD38 + T cells was nearly normal, and the percentage of CD8 +/HLA-DR+ T cells was higher than normal level ( P ＜ 0. 05 ). There was a positive correlation between percentage of CD8+/CD38+ and of CD8 +/HLA-DR+ T cells and viral load ( R = 0. 403, P = 0. 03 for the former and R = 0. 569, P = 0. 09 for the later). Conclusions Effective HAART could decrease immune activation of HIV-infected children significantly. And there was a positive correlation between percentage of CD8 +/CD38 + and of CD8 +/HLA-DR + T cells and viral load, suggesting that the two indicators might be used as the substitution of viral load in resource-limited areas.     

原发性肾病综合征患儿外周血CD4+CD25+调节性T细胞及CD19+CD23+细胞水平的变化及其意义

目的通过观察原发性肾病综合征(PNS)患儿外周血淋巴细胞亚群,尤其是CD4+CD25+调节性T细胞及CD19+CD23+细胞水平的变化,探讨其免疫发病机制。方法采用双标法用流式细胞仪检测25例初发PNS患儿(PNS组)外周血T淋巴细胞亚群(CD3+、CD3+CD4+、CD3+CD8+、CD4+CD25+)、B淋巴细胞亚群(CD3-CD19+、CD19+CD23+)及自然杀伤细胞(CD3-CD16+56+)水平,同时选取同期19例健康儿童作为健康对照组。数据采用SPSS 15.0软件进行统计学分析。结果 PNS组患儿外周血中CD3+、CD3+CD8+、CD4+CD25+、CD19+CD23+淋巴细胞均显著高于健康对照组(Pa<0.05),而自然杀伤细胞则较健康对照组显著降低(P<0.05);PNS组CD3+CD4+、CD4+/CD8+、CD3-CD19+淋巴细胞与健康对照组比较差异无统计学意义。结论体内淋巴细胞亚群的紊乱参与了PNS的发病过程,其中CD4+CD25+调节性T细胞及CD19+CD23+细胞的变化为PNS的免疫治疗目标提供了理论依据。     

Passive smoking alters circulating naïve/memory lymphocyte T‐cell subpopulations in children

Vardavas CI, Plada M, Tzatzarakis M, Marcos A, Warnberg J, Gomez‐Martinez S, Breidenassel C, Gonzalez‐Gross M, Tsatsakis AM, Saris WH., Moreno LA, Kafatos AG. Passive smoking alters circulating naïve/memory lymphocyte T‐cell subpopulations in children.
Pediatr Allergy Immunol 2010: 21: 1171–1178.
© 2010 John Wiley & Sons A/S While it has been indicated that exposure to second‐hand smoke (SHS) can cause a local in vivo response, limited evidence exists on its possible systemic effects from population‐based levels of exposure. We investigated into a possible systemic response in the immune parameters and lymphocyte subsets, i.e. B cell (CD19+), T cell (CD4+CD45RO+, CD4+CD45RA+, CD3+CD45RO+, CD3+CD45RA+) and natural killer (CD3+CD16CD56+) lymphocyte subsets relative to exposure to SHS. Blood was drawn from healthy, verified non‐smoker, adolescent subjects (n = 68, mean age 14.2) and analysed for cotinine, antioxidants and lymphocyte immunophenotyping. SHS exposure was assessed using serum cotinine. Biomarker quantified exposure to SHS was correlated with a linear dose–response reduction in the percentages of memory CD4+CD45RO+ (p = 0.005) and CD3+CD45RO+ T‐cell subsets (p = 0.005 and p = 0.003, respectively) and a linear increase in the percentage of naïve CD4+CD45RA+ and CD3+CD45RA+ T‐cell subsets (p = 0.006 and p = 0.003, respectively). Additionally, higher exposure to SHS was associated with a higher CD4+CD45RA+ count (532 vs. 409 cells/ml, p = 0.017). Moreover, after controlling for age, gender, body mass index and plasma antioxidants, SHS exposure was found to be associated with the percentage of circulating naïve and memory CD4+ and CD3+ T‐cell subpopulations, as revealed through a linear regression analysis. These findings indicate a systemic immunological response in healthy adolescents exposed to population‐based levels of SHS exposure and imply an additional biological pathway for the interaction between exposure to SHS and its adverse effects on human health.     

Foxp3表达与CD+4CD+25调节性T细胞在儿童哮喘发病中的作用

目的研究Foxp3基因表达与CD+4CD+25调节性T细胞在哮喘发病中的作用.方法以确诊哮喘的患儿为研究对象,急性发作期15例、缓解期15例,同期选10例正常儿童作对照,提取外周血单个核细胞(PBMC)进行CD4、CD25表面标志及血浆、培养上清液IL-4、IFN-γ、IL-10和TGF-β等细胞因子的ELISA检测,同时收集哮喘患儿和正常儿童的诱导痰,用RT-PCR方法检测PBMC及诱导痰中转录因子Foxp3-mRNA的表达.结果 PBMC CD+4CD+25T细胞百分率在哮喘急性发作期、缓解期分别为(10.1±2.1)%、(11.7±2.5)%,低于对照组的(15.5±2.7)%(P分别<0.01、<0.05);对照组PBMC在体外培养后CD+4CD+25细胞百分率显著升高,同培养前比较差异有统计学意义(P<0.01).PBMC Foxp3-mRNA表达水平(Foxp3/β-actin) 在哮喘急性发作期、缓解期分别为0.46±0.14 、0.50±0.19,低于对照组0.77±0.22,诱导痰Foxp3-mRNA表达水平哮喘患儿也低于对照组;PBMC在体外培养后对照组Foxp3-mRNA表达水平较培养前升高(P<0.05),而哮喘患儿培养前后Foxp3-mRNA表达水平无显著变化.血浆及培养上清液IFN-γ、TGF-β在哮喘急性发作期、缓解期低于对照组 (P<0.05),且IFN-γ、TGF-β与PBMC Foxp3-mRNA水平、CD+4CD+25细胞百分率呈正相关. 哮喘患儿血浆及培养上清液IL-4显著高于对照组,急性发作期血浆IL-10显著高于对照组,而缓解期与正常组无显著性差异;IL-4、IL-10与PBMC Foxp3-mRNA水平、CD+4CD+25细胞百分率无相关性.结论哮喘患儿的TGF-β分泌不足、Foxp3基因表达降低、CD+4CD+25调节性T细胞数量减少及分化发育障碍可能在儿童哮喘的发病中起重要作用.